Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 30
Filtrar
Más filtros











Intervalo de año de publicación
1.
J Fluoresc ; 2024 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-38805132

RESUMEN

Copper metal is third most abundant trace element in human body. Determination of Cu (II) ions is a burning topic in field of environment protection and food safety because of its significant impact on ecosystem. In this study, 2,6-pyridine dicarboxylic acid (PDA) has been explored as "turn-off" florescent probe for florescent detection of Cu (II) ions. This sensor showed highly selective complexing ability towards Cu (II) ions. Addition of aqueous solution of Cu (II) ions remarkably quenched the fluorescence intensity of PDA while, on contrary, there was no any prominent fluorescence quenching interference on addition of various metal ions. The binding mode of PDA and Cu (II) ions was determined as stoichiometry of 1:1 and it was further confirmed by single crystal XRD analysis. Mechanisms of static and dynamic quenching were confirmed by stern-volmer plot. Limit of detection (LOD) and limit of quantification (LOQ) for Cu (II) ions was calculated as 3.6 µM and 1.23 µM respectively, which is far below the acceptable value (31.5µM) according to the World Health Organization. The use of the sensor for detection of Cu (II) ions in real samples in aqueous media was also performed.

2.
J Fluoresc ; 2024 May 17.
Artículo en Inglés | MEDLINE | ID: mdl-38758342

RESUMEN

Dipicolinic acid (DPA) is a prominent biomarker for Anthrax disease. Bacillus anthracis bacterial endospores is composed of DPA as the significant component, which on over inhalation can cause severe health issues. Such contagious and life-threatening pathogens can be employed as bioweapons or biothreat agents for spreading bioterrorism which is a major risk for national security and public health concerns. Hence, effective detection or a surveillance system is essential for preventing the growth of bioterrorism events. Herein, we have developed a Terbium - 1,10 Phenanthroline (Tb-Phen) based lanthanide luminescence complex with bright green fluorescence. On addition of DPA, the green fluorescence is turn-off at a linear range from 0.6 to 4.762 mM. In this effect, 5D4- 7F5 transition caused by 1,10-phenanthroline to Tb3+ at 544 nm is restricted due to energy transfer quenching and Inner Filter Effect (IFE). The developed probe shows good sensitivity towards the detection of DPA with other coexisting biomolecules and ions with a low Limit of Detection (LOD) of 5.029 µM. The practical feasibility was evaluated in paper strip assay and extended in real samples such as human serum and tap water with satisfactory recovery percentage. Thereby, probe finds promising application in sensing of anthrax spore biomarker (DPA) and biothreat agents.

3.
Microb Cell Fact ; 22(1): 188, 2023 Sep 19.
Artículo en Inglés | MEDLINE | ID: mdl-37726725

RESUMEN

BACKGROUND: Plastics are an indispensable part of our daily life. However, mismanagement at their end-of-life results in severe environmental consequences. The microbial conversion of these polymers into new value-added products offers a promising alternative. In this study, we engineered the soil-bacterium Comamonas testosteroni KF-1, a natural degrader of terephthalic acid, for the conversion of the latter to the high-value product 2-pyrone-4,6-dicarboxylic acid. RESULTS: In order to convert terephthalic acid to 2-pyrone-4,6-dicarboxylic acid, we deleted the native PDC hydrolase and observed only a limited amount of product formation. To test whether this was the result of an inhibition of terephthalic acid uptake by the carbon source for growth (i.e. glycolic acid), the consumption of both carbon sources was monitored in the wild-type strain. Both carbon sources were consumed at the same time, indicating that catabolite repression was not the case. Next, we investigated if the activity of pathway enzymes remained the same in the wild-type and mutant strain. Here again, no statistical differences could be observed. Finally, we hypothesized that the presence of a pmdK variant in the degradation operon could be responsible for the observed phenotype and created a double deletion mutant strain. This newly created strain accumulated PDC to a larger extent and again consumed both carbon sources. The double deletion strain was then used in a bioreactor experiment, leading to the accumulation of 6.5 g/L of product in 24 h with an overall productivity of 0.27 g/L/h. CONCLUSIONS: This study shows the production of the chemical building block 2-pyrone-4,6-dicarboxylic acid from terephthalic acid through an engineered C. testosteroni KF-1 strain. It was observed that both a deletion of the native PDC hydrolase as well as a pmdK variant is needed to achieve high conversion yields. A product titer of 6.5 g/L in 24 h with an overall productivity of 0.27 g/L/h was achieved.


Asunto(s)
Comamonas testosteroni , Comamonas testosteroni/genética , Carbono , Ácidos Dicarboxílicos , Hidrolasas
4.
Bioresour Technol ; 377: 128956, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-36965585

RESUMEN

2-Pyrone-4,6-dicarboxylic acid (PDC) is a valuable building block molecule produced from lignin-derived aromatic compounds by biological funneling. This study aimed to design a fermentation process for producing PDC from vanillic acid, which could be applied at an industrial production. Metabolomic analysis revealed that a high primary metabolic activity within cells was required to improve the production efficiency. Moreover, a medium with ammonium salts and no alkali metals was advantageous because it suppressed the formation of PDC-metal complexes. Resulting optimized process yielded the highest PDC titer and productivity ever reported: 99.9 g/L and 1.69 g/L/h, respectively. Per batch, 190 g of PDC was produced per liter of initial culture media, and the final liquid volume was 1.9 L. This study demonstrates the design of fermentation processes for the advanced industrial utilization of lignin by biological funneling.


Asunto(s)
Lignina , Ácido Vanílico , Lignina/metabolismo , Fermentación , Ácidos Dicarboxílicos
5.
J Fluoresc ; 33(3): 1057-1065, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-36565411

RESUMEN

Tb3+ luminescence is enhanced by complex formation in aqueous phases as its pyridine 2,6 dicarboxylate (dpa2-) complexes by using experimental spectroscopic techniques and theoretical time-dependent density functional theory (TD-DFT) calculations. The fluorimetric titration of Tb3+ ion with dpa2- ion is followed at λext/λems = 310/490 nm and 310/545 nm, the emission intensities of which are graphed against the mol ratios of the ligand to metal ion [moles of dpa2-/mol of Tb3+]. Experimental results denote that the tris complex; [Tb(dpa)3]3- is the most stable form at pH > 5.3. Molecular absorption spectra of tris complex shows a batho-chromic shift of 222 nm of dpa2- band to 232 nm accompanied by the hyper-chromic effect at 272 nm band. The luminescence intensities at 490, 545, 592 and 620 nm are enhanced over 100 times in tris complex. The coordination of complexes calculated by thermodynamic cycles and with supporting the experimental result, the most stable form was found to be nine coordinated tris complex; [Tb(dpa)]3-. The theoretical TD-DFT calculations perfectly matched the experimental absorption and emission bands of tris-complex. The novelty of this study is to present the first theoretical calculation of the phosphorescence results and energy transfer process for emission path of Tb3+ and pda2- aqua complexes.

6.
Bioresour Technol ; 363: 127836, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36031121

RESUMEN

In grass biomass, hydroxycinnamic acids (HCAs) play crucial roles in the crosslinking of lignin and polysaccharides and can be easily extracted by mild alkaline pretreatment, albeit heterogeneously. Here, HCAs were extracted from bamboo and rice straw as model grass biomass with different HCAs composition, and microbial funneling was then conducted to produce 2-pyrone-4,6-dicarboxylic acid (PDC) and (4S)-3-carboxymuconolactone (4S-3CML), promising building blocks for bio-based polymers, respectively. Pseudomonas putida PpY1100 engineered for efficient microbial funneling completely converted HCAs to PDC and 4S-3CML with high titers of 3.9-9.3 g/L and molar yields of 92-99%, respectively. The enzymatic saccharification efficiencies of lignocellulose after HCAs extraction were 29.5% in bamboo and 73.8% in rice straw, which are 8.9 and 6.8 times higher than in alkaline-untreated media, respectively. These results provide a green-like process for total valorization of grass biomass through enzymatic saccharification integrated with upgrading heterogeneous HCAs to a valuable single chemical via microbial funneling.


Asunto(s)
Lignina , Oryza , Biomasa , Ácidos Cumáricos , Hidrólisis , Lignina/química , Oryza/química , Poaceae , Polisacáridos/química
7.
Bioresour Technol ; 352: 127106, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-35378283

RESUMEN

Lignin valorization depends on microbial upcycling of various aromatic compounds in the form of a complex mixture, including p-coumaric acid and ferulic acid. In this study, an engineered Pseudomonas putida strain utilizing lignin-derived monomeric compounds via biological funneling was developed to produce 2-pyrone-4,6-dicarboxylic acid (PDC), which has been considered a promising building block for bioplastics. The biosynthetic pathway for PDC production was established by introducing the heterologous ligABC genes under the promoter Ptac in a strain lacking pcaGH genes to accumulate a precursor of PDC, i.e., protocatechuic acid. Based on the culture optimization, fed-batch fermentation of the final strain resulted in 22.7 g/L PDC with a molar yield of 1.0 mol/mol and productivity of 0.21 g/L/h. Subsequent purification of PDC at high purity was successfully implemented, which was consequently applied for the novel polyester.


Asunto(s)
Pseudomonas putida , Ácidos Dicarboxílicos/metabolismo , Lignina/metabolismo , Poliésteres/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Pironas
8.
Anal Bioanal Chem ; 414(8): 2767-2774, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35106614

RESUMEN

Analysis of the dipicolinic acid (DPA) released from Clostridium botulinum spores during thermal processing is crucial to obtaining a mechanistic understanding of the factors involved in spore heat resistance and related food safety applications. Here, we developed a novel mixed-mode liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detection of the DPA released from C. botulinum type A, nonproteolytic types B and F strains, and nonpathogenic surrogate Clostridium sporogenes PA3679 spores. DPA was retained on a mixed-mode C18/anion exchange column and was detected using electrospray ionization (ESI) positive mode within a 4-min analysis time. The intraday and interday precision (%CV) was 1.94-3.46% and 4.04-8.28%, respectively. Matrix effects were minimal across proteolytic type A Giorgio-A, nonproteolytic types QC-B and 202-F, and C. sporogenes PA3679 spore suspensions (90.1-114% of spiked DPA concentrations). DPA recovery in carrot juice and beef broth ranged from 105 to 118%, indicating limited matrix effects of these food products. Experiments that assessed the DPA released from Giorgio-A spores over the course of a 5-min thermal treatment at 108 °C found a significant correlation (R = 0.907; P < 0.05) between the log reduction of spores and amount of DPA released. This mixed-mode LC-MS/MS method provides a means for rapid detection of DPA released from C. botulinum spores during thermal processing and has the potential to be used for experiments in the field of food safety that assess the thermal resistance characteristics of various C. botulinum spore types.


Asunto(s)
Clostridium botulinum , Ácidos Picolínicos , Cromatografía Liquida , Clostridium botulinum/química , Calor , Ácidos Picolínicos/análisis , Esporas Bacterianas/química , Espectrometría de Masas en Tándem
9.
Metab Eng ; 66: 148-156, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33895365

RESUMEN

2-Pyrone-4,6-dicarboxylic acid (PDC), a chemically stable intermediate that naturally occurs during microbial degradation of lignin by bacteria, represents a promising building block for diverse biomaterials and polyesters such as biodegradable plastics. The lack of a chemical synthesis method has hindered large-scale utilization of PDC and metabolic engineering approaches for its biosynthesis have recently emerged. In this study, we demonstrate a strategy for the production of PDC via manipulation of the shikimate pathway using plants as green factories. In tobacco leaves, we first showed that transient expression of bacterial feedback-resistant 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase (AroG) and 3-dehydroshikimate dehydratase (QsuB) produced high titers of protocatechuate (PCA), which was in turn efficiently converted into PDC upon co-expression of PCA 4,5-dioxygenase (PmdAB) and 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase (PmdC) derived from Comamonas testosteroni. We validated that stable expression of AroG in Arabidopsis in a genetic background containing the QsuB gene enhanced PCA content in plant biomass, presumably via an increase of the carbon flux through the shikimate pathway. Further, introducing AroG and the PDC biosynthetic genes (PmdA, PmdB, and PmdC) into the Arabidopsis QsuB background, or introducing the five genes (AroG, QsuB, PmdA, PmdB, and PmdC) stacked on a single construct into wild-type plants, resulted in PDC titers of ~1% and ~3% dry weight in plant biomass, respectively. Consistent with previous studies of plants expressing QsuB, all PDC producing lines showed strong reduction in lignin content in stems. This low lignin trait was accompanied with improvements of biomass saccharification efficiency due to reduced cell wall recalcitrance to enzymatic degradation. Importantly, most transgenic lines showed no reduction in biomass yields. Therefore, we conclude that engineering plants with the proposed de-novo PDC pathway provides an avenue to enrich biomass with a value-added co-product while simultaneously improving biomass quality for the supply of fermentable sugars. Implementing this strategy into bioenergy crops has the potential to support existing microbial fermentation approaches that exploit lignocellulosic biomass feedstocks for PDC production.


Asunto(s)
Arabidopsis , Poliésteres , Arabidopsis/genética , Biomasa , Lignina , Pironas
10.
Metab Eng ; 65: 111-122, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33741529

RESUMEN

Valorization of lignin, an abundant component of plant cell walls, is critical to enabling the lignocellulosic bioeconomy. Biological funneling using microbial biocatalysts has emerged as an attractive approach to convert complex mixtures of lignin depolymerization products to value-added compounds. Ideally, biocatalysts would convert aromatic compounds derived from the three canonical types of lignin: syringyl (S), guaiacyl (G), and p-hydroxyphenyl (H). Pseudomonas putida KT2440 (hereafter KT2440) has been developed as a biocatalyst owing in part to its native catabolic capabilities but is not known to catabolize S-type lignin-derived compounds. Here, we demonstrate that syringate, a common S-type lignin-derived compound, is utilized by KT2440 only in the presence of another energy source or when vanAB was overexpressed, as syringate was found to be O-demethylated to gallate by VanAB, a two-component monooxygenase, and further catabolized via extradiol cleavage. Unexpectedly, the specificity (kcat/KM) of VanAB for syringate was within 25% that for vanillate and O-demethylation of both substrates was well-coupled to O2 consumption. However, the native KT2440 gallate-cleaving dioxygenase, GalA, was potently inactivated by 3-O-methylgallate. To engineer a biocatalyst to simultaneously convert S-, G-, and H-type monomers, we therefore employed VanAB from Pseudomonas sp. HR199, which has lower activity for 3MGA, and LigAB, an extradiol dioxygenase able to cleave protocatechuate and 3-O-methylgallate. This strain converted 93% of a mixture of lignin monomers to 2-pyrone-4,6-dicarboxylate, a promising bio-based chemical. Overall, this study elucidates a native pathway in KT2440 for catabolizing S-type lignin-derived compounds and demonstrates the potential of this robust chassis for lignin valorization.


Asunto(s)
Pseudomonas putida , Lignina , Pseudomonas putida/genética , Pironas
11.
Acta Crystallogr C Struct Chem ; 76(Pt 12): 1076-1084, 2020 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-33273145

RESUMEN

The FeIII ion as a ubiquitous metal plays a key role in biochemical processes. Iron deficiency or excess in the human body can induce various diseases. Thus, effective detection of the FeIII ion has been deemed an issue of focus. To develop more crystalline chemical sensors for the selective detection of Fe3+, two novel two-dimensional (2D) coordination polymers, namely, poly[[[µ-bis(pyridin-4-yl)amine-κ2N:N'](µ-naphthalene-2,6-dicarboxylato-κ2O2:O6)zinc(II)] 0.5-hydrate], {[Zn(C12H6O4)(C10H9N3)]·0.5H2O}n, 1, and poly[(4,4'-dimethyl-2,2'-bipyridine-κ2N,N')(µ-naphthalene-2,6-dicarboxylato-κ2O2:O6)hemi(µ-naphthalene-2,6-dicarboxylic acid-κ2O2:O6)copper(II)] [Cu(C12H6O4)(C12H12N2)(C12H8O4)0.5]n, 2, have been prepared using solvothermal methods. Single-crystal X-ray diffraction analysis shows that compound 1 is an undulating twofold interpenetrated 2D (4,4)-sql network and compound 2 is a twofold interpenetrated 2D honeycomb-type network with a (6,3)-hcb topology. In addition, 1 exhibits highly selective sensing for the Fe3+ ion.

12.
Front Microbiol ; 11: 1841, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32849439

RESUMEN

Pseudomonas aeruginosa isolated from the plant rhizosphere has been widely used as an effective strain in biological control against plant disease. This bacterium promotes plant growth and protect plants against various phytopathogens through the production of phenazine metabolites. In this study, the strain P. aeruginosa Y12 with anti-Beauveria bassiana activity was isolated from the gut of housefly larvae. It was comparatively analyzed with the strain P. aeruginosa P18, which showed no anti-B. bassiana activity. Genomic and metabolomic methods were used to obtain a comprehensive understanding of the antimicrobial mechanism of Y12. After whole-genome resequencing of the two strains, a total of 7,087 non-synonymous single-nucleotide polymorphisms (nsSNPs), 1079 insertions and deletions (InDels), 62 copy-number variations (CNVs) and 42 structural variations (SV) were found in both strains. We analyzed the differentially abundant metabolites between Y12 and P18, and identified six bioactive compounds that could be associated with the antimicrobial activity of Y12. Additionally, we found that, unlike other previously reported rhizospheric P. aeruginosa strains, Y12 could produce both phenazine-1,6-dicarboxylic acid (PDC) and pyocyanin (PYO) at significantly higher concentrations than P18. As B. bassiana is an effective biological insecticide that can cause high mortality in adult houseflies but has little effect on housefly larvae, we believe that P. aeruginosa Y12, identified in housefly larvae but not in adults, were beneficial for the development of housefly larvae and could protect them from B. bassiana infection through the production of toxic metabolites.

13.
J Biosci Bioeng ; 130(1): 71-75, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32238321

RESUMEN

Lignosulfonate is a by-product of the cooking process by sulfite pulping for paper manufacturing. The treatment of wood chips by various salts of sulfurous acid solubilizes lignin to produce a cellulose-rich wood pulp. Developing a technique for the conversion of lignosulfonate by-product to high value materials has an important industrial utility. Sphingobium sp. strain SYK-6, which was isolated from pulping wastewater, is one of the best enzymatically or genetically characterized bacteria for degrading lignin-derived aromatics. We have previously established a system for the production of 2-pyrone-4,6-dicarboxylic acid (PDC), a novel platform chemical that can produce a variety of bio-based polymers, by introducing of ligA, ligB, and ligC genes from SYK-6 into a mutant strain of Pseudomonas putida PpY1100. In this study, extracts from lignosulfonates, which were desulphonated and depolymerized by alkaline oxidation, were evaluated as substrates for microbiological conversion to PDC by the transgenic bacteria.


Asunto(s)
Lignina/metabolismo , Extractos Vegetales/metabolismo , Pseudomonas putida/metabolismo , Pironas/metabolismo , Sphingomonadaceae/metabolismo , Celulosa/metabolismo , Ácidos Dicarboxílicos/metabolismo , Pseudomonas putida/genética , Sphingomonadaceae/genética , Residuos/análisis
14.
J Agric Food Chem ; 68(8): 2373-2380, 2020 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-32013409

RESUMEN

Pseudomonas chlororaphis have been demonstrated to be environmentally friendly biocontrol strains, and most of them can produce phenazine compounds. Phenazine-1,6-dicarboxylic acid (PDC), with a potential antibacterial activity, is generally found in Streptomyces but not in Pseudomonas. The present study aimed to explore the feasibility of PDC synthesis and the function of PhzG in Pseudomonas. A PDC producer was constructed by replacing phzG in P. chlororaphis with lphzG from Streptomyces lomondensis. Through gene deletion, common start codon changing, gene silence, and in vitro assay, our result revealed that the yield of PDC in P. chlororaphis is associated with the relative expression of phzG to phzA and phzB. In addition, it is found that PDC can be spontaneously synthesized without PhzG. This study provides an efficient way for PDC production and promotes a better understanding of PhzG function in PDC biosynthesis. Moreover, this study gives an alternative opportunity for developing new antibacterial biopesticides.


Asunto(s)
Antibacterianos/biosíntesis , Proteínas Bacterianas/metabolismo , Fenazinas/metabolismo , Pseudomonas chlororaphis/genética , Pseudomonas chlororaphis/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Eliminación de Gen , Datos de Secuencia Molecular , Pseudomonas chlororaphis/enzimología , Alineación de Secuencia
15.
Acta Crystallogr C Struct Chem ; 75(Pt 6): 723-727, 2019 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-31166925

RESUMEN

The design and synthesis of 3d-4f heterometallic coordination polymers have attracted much interest due to the intriguing diversity of their architectures and topologies. Pyridine-2,6-dicarboxylic acid (H2pydc) has a versatile coordination mode and has been used to construct multinuclear and heterometallic compounds. Two isostructural centrosymmetric 3d-4f coordination compounds constructed from pyridine-2,6-dicarboxylic acid and 4,4'-bipyridine (bpy), namely 4,4'-bipyridine-1,1'-diium diaquabis(µ2-pyridine-2,6-dicarboxylato)tetrakis(pyridine-2,6-dicarboxylato)bis[4-(pyridin-4-yl)pyridinium]cobalt(II)dieuropium(III) octahydrate, (C10H10N2)[CoEu2(C10H9N2)2(C7H3NO4)6(H2O)2]·8H2O, (I), and 4,4'-bipyridine-1,1'-diium diaquabis(µ2-pyridine-2,6-dicarboxylato)tetrakis(pyridine-2,6-dicarboxylato)bis[4-(pyridin-4-yl)pyridinium]cobalt(II)diterbium(III) octahydrate, (C10H10N2)[CoTb2(C10H9N2)2(C7H3NO4)6(H2O)2]·8H2O, (II), were synthesized under hydrothermal conditions and characterized by IR and fluorescence spectroscopy, thermogravimetric analysis and powder X-ray diffraction. Both compounds crystallize in the triclinic space group P-1. The EuIII and TbIII cations adopt nine-coordinated distorted tricapped trigonal-prismatic geometries bridged by three pydc2- ligands. The CoII cation has a six-coordination environment formed by two pydc2- ligands, two bpy ligands and two coordinated water molecules. Adjacent molecules are connected by π-π stacking interactions to form a one-dimensional chain, which is further extended into a three-dimensional supramolecular network by multipoint hydrogen bonds.

16.
Biomolecules ; 9(5)2019 05 16.
Artículo en Inglés | MEDLINE | ID: mdl-31100934

RESUMEN

4-oxo-4H-pyran-2.6-dicarboxylic acid (chelidonic acid, ChA) in the native state and in the complex with calcium [Ca(ChA)(H2O)3], named saucalchelin (CaChA), was isolated from the extract of Saussurea controversa leaves for the first time for the Asteraceae family. The structure of ChA was determined by NMR, MS and confirmed by X-ray analysis of its monomethyl ester, and CaChA was described by IR, ICP-MS, CHN analysis. The yield of ChA and CaChA was 45 mg/g and 70 mg/g of extract, respectively. The osteogenic activity of ChA, n-monobutyl ester of chelidonic acid, and CaChA has been studied in vitro in a 21-day culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) in a standard nutrient medium without osteogenic supplements. CaChA significantly stimulated the growth of cell mass and differentiation of hAMMSCs into osteoblasts with subsequent mineralization of the culture and it may be a promising substance for accelerating bone tissue regeneration and engineering.


Asunto(s)
Diferenciación Celular , Células Madre Mesenquimatosas/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Extractos Vegetales/farmacología , Piranos/farmacología , Saussurea/química , Células Cultivadas , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/citología , Osteoblastos/metabolismo , Extractos Vegetales/química , Piranos/química
17.
J Oleo Sci ; 67(9): 1059-1070, 2018 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-30111682

RESUMEN

In this review, we show novel methods for utilizing lignocellulosic biomass, polysaccharides, and lignin. Firstly, the simultaneous enzymatic saccharification and comminution (SESC) of plant materials is described as an extraction method for lignocellulosic biomass that does not require toxic reagents or organic solvents. Secondly, we demonstrate the material utilization of non-deteriorated lignocellulosic biomass extracted by SESC, such as for sugar and ethanol synthesis, and as a heatproof filler. Finally, we exhibit the use of a functional monomer (e.g., in disinfection chemicals, cesium chelation, and building blocks for polymers), 2-pyrone-4,6-dicarboxylic acid, derived from lignin via metabolic degradation.


Asunto(s)
Biomasa , Biotecnología/métodos , Lignina , Polisacáridos , Extracción en Fase Sólida/métodos , Etanol , Pironas , Azúcares
18.
ACS Synth Biol ; 7(9): 2296-2307, 2018 09 21.
Artículo en Inglés | MEDLINE | ID: mdl-30096230

RESUMEN

2-Pyrone-4,6-dicarboxylic acid (PDC) is a pseudoaromatic dicarboxylic acid and is a promising biobased building block chemical that can be used to make diverse polyesters with novel functionalities. In this study, Escherichia coli was metabolically engineered to produce PDC from glucose. First, an efficient biosynthetic pathway for PDC production from glucose was suggested by in silico metabolic flux simulation. This best pathway employs a single-step biosynthetic route to protocatechuic acid (PCA), a metabolic precursor for PDC biosynthesis. On the basis of the selected PDC biosynthetic pathway, a shikimate dehydrogenase (encoded by aroE)-deficient E. coli strain was engineered by introducing heterologous genes of different microbial origin encoding enzymes responsible for converting 3-dehydroshikimate (DHS) to PDC, which allowed de novo biosynthesis of PDC from glucose. Next, production of PDC was further improved by applying stepwise rational metabolic engineering strategies. These include elimination of feedback inhibition on 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase (encoded by aroG) by overexpressing a feedback-resistant variant, enhancement of the precursor phosphoenolpyruvate supply by changing the native promoter of the ppsA gene with the strong trc promoter, and reducing accumulation of the major byproduct DHS by overexpression of a DHS importer (encoded by shiA). Furthermore, cofactor (NADP+/NADPH) utilization was manipulated through genetic modifications of the E. coli soluble pyridine nucleotide transhydrogenase (encoded by sthA), and the resultant impact on PDC production was investigated. Fed-batch fermentation of the final engineered E. coli strain allowed production of 16.72 g/L of PDC from glucose with the yield and productivity of 0.201 g/g and 0.172 g/L/h, respectively, representing the highest PDC production performance indices reported to date.


Asunto(s)
Escherichia coli/metabolismo , Glucosa/metabolismo , Ingeniería Metabólica , Pironas/metabolismo , Técnicas de Cultivo Celular por Lotes , Vías Biosintéticas , Dioxigenasas/genética , Dioxigenasas/metabolismo , Hidroliasas/genética , Hidroliasas/metabolismo , NADP/química , NADP Transhidrogenasas/genética , NADP Transhidrogenasas/metabolismo , Plásmidos/genética , Plásmidos/metabolismo , Ácido Shikímico/metabolismo
19.
Ultrason Sonochem ; 46: 68-78, 2018 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-29739514

RESUMEN

A self-assembly of pyridine-2,6-dicarboxylate with Cu(II) and Mn(II) under ultrasonic and microwave irradiation gave the two coordination polymers [Cu(PDA)(H2O)1.5]n (1) and [Mn(PDA)(H2O)1.5]n (2). Their structures were characterized using IR, elemental analysis, X-ray diffraction (XRD) and spectroscopic methods. The corresponding α-Mn3O4 and CuO nanoparticles were synthesized by calcination of 1 and 2 in air at 600 °C. Transmission electron microscopy (TEM) reveals a sphere-like morphology for the Mn3O4 nanoparticles. Shrinkage of the particle size from 90 nm (by conventional synthesis of the precursor) to 19 nm (ultrasonic-assisted) takes place, indicating the great effect of ultrasonication. CuO nanoparticles were of semispherical (conventional and ultrasonic-assisted methods) and hexagonal shapes (microwave irradiation) with an average diameter of 7, 15 and 25 nm, respectively. The catalytic performance of the coordination polymers towards degradation of methylene blue and methyl orange in the presence of hydrogen peroxide was studied. Using the same dose, catalyst 1 proved to be more efficient in color removal of both MB and MO than catalyst 2 did. Recycling test for 2 showed that it is a recyclable catalyst with no structural changes over three recycling experiments.

20.
J Biosci Bioeng ; 125(6): 717-722, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29395960

RESUMEN

Actual biomass of microalgae was tested as a fermentation substrate for microbial production of 2-pyrone 4,6-dicarboxylic acid (PDC). Acid-hydrolyzed green microalgae Chlorella emersonii (algae hydrolysate) was diluted to adjust the glucose concentration to 2 g/L and supplemented with the nutrients of Luria-Bertani (LB) medium (tryptone 10 g/L and yeast extract 5 g/L). When the algae hydrolysate was used as a fermentation source for recombinant Escherichia coli producing PDC, 0.43 g/L PDC was produced with a yield of 20.1% (mol PDC/mol glucose), whereas 0.19 g/L PDC was produced with a yield of 8.6% when LB medium supplemented with glucose was used. To evaluate the potential of algae hydrolysate alone as a fermentation medium for E. coli growth and PDC production, the nutrients of LB medium were reduced from the algae hydrolysate medium. Interestingly, 0.17 g/L PDC was produced even without additional nutrient, which was comparable to the case using pure glucose medium with nutrients of LB medium. When using a high concentration of hydrolysate without additional nutrients, 1.22 g/L PDC was produced after a 24-h cultivation with the yield of 16.1%. Overall, C. emersonii has high potential as cost-effective fermentation substrate for the microbial production of PDC.


Asunto(s)
Chlorella/metabolismo , Escherichia coli/metabolismo , Fermentación , Microalgas/metabolismo , Pironas/metabolismo , Biomasa , Hidrolasas de Éster Carboxílico/metabolismo , Chlorella/enzimología , Chlorella/crecimiento & desarrollo , Escherichia coli/genética , Glucosa/metabolismo , Hidrólisis , Microalgas/enzimología , Microalgas/crecimiento & desarrollo , Organismos Modificados Genéticamente
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA